TW201902868A - Methods of synthesizing an mcl-1 inhibitor - Google Patents

Abstract

Disclosed are intermediates and methods of synthesizing Compound 1.

Description

Method for synthesizing MCL-1 inhibitor

Bone marrow cell leukemia 1 (Mcl-1) is an important anti-apoptotic member of the BCL-2 family of proteins and a major regulator of cell survival. Amplification of the MCL1 gene and / or overexpression of the Mcl-1 protein has been observed in a variety of cancer types, and is often involved in tumor development. In fact, MCL1 is one of the most frequently amplified genes in human cancers. In many malignancies, Mcl-1 is a key survival factor and has been shown to mediate resistance to multiple anticancer agents.

Mcl-1 promotes cell survival by binding to pro-apoptotic proteins such as Bim, Noxa, Bak, and Bax and neutralizing its death-inducing activity. This inhibition of Mcl-1 releases these pro-apoptotic proteins, which usually results in induction of apoptosis in tumor cells that depend on Mcl-1 for survival. Therefore, promising strategies for targeting the Mcl-1 line, alone or in combination with other therapies, to treat numerous malignancies and overcome drug resistance in many human cancers. The chemical name of this compound is ( R _a )-(+)-17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13 , 22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] Thirty-eight carbon-1 (37), 4 (38) , 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-carboxylic acid (referred to as compound 1): Compound 1 is a potent Mcl-1 inhibitor, as described in more detail below. Therefore, there is a need to develop a new method for synthesizing Compound 1 in an efficient manner.

Methods and intermediates useful for the synthesis of compound 1 are provided herein: Compound 1

In some embodiments, the following intermediates and any salts thereof, and their synthesis are disclosed:

In some embodiments, it provided herein for synthesizing _{(R a) - (+)} - 17- -5,13,14,22- chloro-28-oxa-2,9-four dithia - 5,6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35-Tridecene-23-carboxylic acid (Compound 1): Compound 1

As shown in Example 1 below, Compound 1 is a potent Mcl-1 inhibitor and can be used to treat cancer including malignant hematological diseases such as acute myeloid leukemia, multiple myeloma, mantle cell lymphoma, and chronic lymphocytic leukemia , Diffuse large B-cell lymphoma, Burkitt's lymphoma, follicular lymphoma, and solid tumors such as non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), breast cancer, neuroblastoma, prostate cancer, Melanoma, pancreatic cancer, uterine cancer, endometrial cancer, and colon cancer. Due to the structural complexity of Compound 1, achieving effective synthesis can be an important aspect in developing Compound 1 as a potential cancer treatment. Intermediate

In some embodiments, (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydradino) adipate (Intermediate 1) is disclosed: Intermediate 1

In some embodiments, a compound of formula (d) is disclosed: (d) wherein R1 is a protecting group or hydrogen. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (a) is (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid (Intermediate 6): Intermediate 6

In some embodiments, compounds of formula (e) are disclosed: (e) wherein R1 is a protecting group or hydrogen. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound is having the formula (b) of (R _a) -3- (2- carboxyethyl) -6-chloro-7- (3 - (((4-methoxybenzyl) oxy (Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitro Phenyl) ethylamine (1: 1 salt) (Intermediate 7): Intermediate 7

In some embodiments, 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester (Intermediate 12) is disclosed: Intermediate 12

In some embodiments, 3-((((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol (Intermediate 13 ): Intermediate 13 In some embodiments, 3-((((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol ( Intermediate 14): Intermediate 14

In some embodiments, 3-((((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate ( Intermediate 15): Intermediate 15

In some embodiments, 3-((((3-((acetamidothio) methyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1- Glycolate (Intermediate 16): Intermediate 16 . synthesis

In some embodiments, a method for synthesizing (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydradino) adipate (Intermediate 1) is disclosed: Intermediate 1 includes the following steps: (i) contacting 2-bromo-3-chloroaniline and methyl 2- pendant cyclopentane-1-carboxylic acid with a diazotizing agent in an acidic aqueous system; (ii) bringing Methyl 2- pendant cyclopentane-1-carboxylic acid and an aqueous base are added to the acidic aqueous system; (iii) the separated plutonium; (iv) the plutonium is contacted with an acidic solution; and (v) separated (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydradino) adipate (Intermediate 1). In some embodiments, the diazotizing agent is NaNO ₂ , Ca (NO ₂ ) ₂ or KNO ₂ . In some embodiments, the diazotizing agent is NaNO ₂ . In some embodiments, the acidic aqueous system comprises a protic acid and water. In some embodiments, the protonic acid is hydrochloric acid. In some embodiments, the aqueous base is potassium acetate, potassium hydroxide, sodium hydroxide, potassium carbonate, sodium carbonate, or potassium phosphate. In some embodiments, the aqueous base is potassium acetate. In some embodiments, the acidic solution comprises an acid and an alcohol. In some embodiments, the acidic solution comprises concentrated sulfuric acid and methanol, methanesulfonic acid and methanol or p-toluenesulfonic acid and methanol. In some embodiments, the acidic solution comprises concentrated sulfuric acid and methanol.

In some embodiments, the synthesis of methyl 7-bromo-6-chloro-3- (3-methoxy-3- pendoxypropyl) -1H-indole-2-carboxylic acid (Intermediate 2) is disclosed Methods: Intermediate 2 includes the following steps: (i) (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydradino) adipate (Intermediate 1) and Contact with an acidic solution; and (ii) isolating methyl 7-bromo-6-chloro-3- (3-methoxy-3- pendoxypropyl) -1H-indole-2-carboxylic acid (intermediate 2) . In some embodiments, the acidic solution comprises an acid and an alcohol. In some embodiments, the acidic solution comprises concentrated sulfuric acid and methanol, methanesulfonic acid and methanol or p-toluenesulfonic acid and methanol. In some embodiments, the acidic solution comprises concentrated sulfuric acid and methanol. In some embodiments, the method of synthesizing intermediate 2 further includes the step of heating the acidic solution before isolating intermediate 2.

In some embodiments, a method for synthesizing a compound of formula (a) is disclosed: (a) wherein R1 is a protecting group or hydrogen, including the following steps: (i) contacting 1,5-dimethyl-1H-pyrazole-3-carboxylic acid ethyl ester with a reducing agent in the presence of a first solvent to Forming a first solution; (ii) isolating (1,5-dimethyl-1H-pyrazol-3-yl) methanol; (iii) making (1,5-dimethyl-1H) in the presence of a second solvent -Pyrazol-3-yl) methanol is contacted with a brominating agent to form (4-bromo-1,5-dimethyl-1H-pyrazol-3-yl) methanol; (iv) separating (4-bromo-1 , 5-dimethyl-1H-pyrazol-3-yl) methanol; (v) (4-bromo-1,5-dimethyl-1H-pyrazol-3-yl) methanol in a third solvent Contact with a base (optionally a phase transfer catalyst) and a protecting group precursor; and (v) separating a compound of formula (a). In some embodiments, the reducing agent is selected from the group consisting of lithium aluminum hydride (LAH), diisobutyl aluminum hydride (DIBAL), lithium borohydride (LiBH ₄ ), sodium bis (2-methoxyethoxy) aluminum hydride ( Red-Al®) and sodium borohydride (NaBH ₄ ). In some embodiments, the reducing agent is lithium aluminum hydride. In some embodiments, the first solvent is selected from the group consisting of toluene, THF, 2-methyltetrahydrofuran, MTBE, methanol, ethanol, and ether. In some embodiments, the first solvent is tetrahydrofuran. In some embodiments, the second solvent is tetrahydrofuran. In some embodiments, the brominating agent is 1,3-dibromo-5,5-dimethylhydantoin (DBDMH). In some embodiments, the brominating agent is N-bromosuccinimide. In some embodiments, the base is lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride, lithium hydride, or potassium hydride. In some embodiments, a phase transfer catalyst is used, such as Bu ₄ N · HSO ₄ or benzyltrimethylammonium chloride. In some embodiments, the base is potassium hydroxide and the phase transfer catalyst is tetrabutylammonium hydrogen sulfate. In some embodiments, the protecting group precursor is 1- (chloromethyl) -4-methoxybenzene. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (a) is 4-bromo-3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole (Intermediate 3): Intermediate 3

In some embodiments, a method for synthesizing a compound of formula (b) is disclosed: (B) wherein R1 is a protecting group or hydrogen, including the following steps: (i) contacting a compound of formula (a) with a metallizing agent in the presence of a solvent; (ii) adding 2-isopropoxy- 4,4,5,5-tetramethyl-1,3,2-dioxaborolane; (iii) adding an acidic solvent solution; and (iv) isolating the compound of formula (b). In some embodiments, the metallizing agent is n-butyllithium. In some embodiments, the solvent is tetrahydrofuran. In some embodiments, the acidic solvent solution comprises acetic acid and toluene. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (a) is 4-bromo-3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole (Intermediate 3). In some embodiments, the compound of formula (b) is 3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-4- (4,4,5 , 5-tetramethyl-1,3,2-dioxolane-2-yl) -1H-pyrazole (Intermediate 4): Intermediate 4

In some embodiments, a method for synthesizing a compound of formula (c) is disclosed: (c) wherein R1 is a protecting group or hydrogen, including the following steps: (i) combining a compound of formula (b) with 7-bromo-6-chloro-3- (3-methoxy-3- pendantoxy (Propyl) -1H-indole-2-carboxylic acid methyl ester (Intermediate 2) is contacted with a palladium catalyst in the presence of a base and a solvent; and (ii) the compound of formula (c) is isolated. In some embodiments, the palladium catalyst is selected from tetrakis (triphenylphosphine) palladium (0), bis (di-tertiarybutyl (4-dimethylaminophenyl) phosphine) palladium (II) dichloride, 1,1'-bis (tertiary-butylphosphine) ferrocene palladium dichloride. In some embodiments, the base is potassium carbonate or potassium phosphate. In some embodiments, the palladium catalyst is 1,1′-bis (tertiary-butylphosphine) ferrocene palladium dichloride. In some embodiments, the solvent is difluorene, water, or a combination thereof. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (b) is 3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-4- (4,4,5 , 5-tetramethyl-1,3,2-dioxolane-2-yl) -1H-pyrazole (Intermediate 4). In some embodiments, the compound of formula (c) is (±) -methyl 6-chloro-3- (3-methoxy-3-sideoxypropyl) -7- (3-((( 4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1H-indole-2-formate (Intermediate 5): Intermediate 5

In some embodiments, a method for synthesizing a compound of formula (d) is disclosed: (d) where R1 is a protecting group or hydrogen, including the following steps: (i) contacting a compound of formula (c) with a methylating agent in a solvent; (ii) methylation obtained by treatment with an ester hydrolysis reagent Derivatives, and (iii) isolate compounds of formula (d). In some embodiments, the compound of Formula (c) is Intermediate 5. In some embodiments, the ester hydrolysis reagent is a hydroxide base selected from the group consisting of lithium hydroxide, potassium hydroxide, and sodium hydroxide. In some embodiments, the methylating agent is selected from the group consisting of methyl iodide, dimethyl sulfate, and dimethylformamide-dimethylacetal (DMF-DMA). In some embodiments, the methylation reagent is dimethylformamide-dimethylacetal. In some embodiments, the solvent is toluene.

In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (c) is (±) -methyl 6-chloro-3- (3-methoxy-3-sideoxypropyl) -7- (3-((( 4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1H-indole-2-formate (Intermediate 5). In some embodiments, the compound of formula (d) is (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid (Intermediate 6): Intermediate 6

In some embodiments, a method for synthesizing a compound of formula (e) is disclosed: (e) wherein R1 is a protecting group or hydrogen, and includes the following steps: (i) combining a compound of formula (d) with (1 R ) -1- (2-nitrophenyl) ethylamine hydrochloride in a base Contact with a solvent; and (ii) isolating a compound of formula (e). In some embodiments, the base is sodium hydroxide, potassium hydroxide, diisopropylethylamine, or triethylamine. In some embodiments, the base is sodium hydroxide. In some embodiments, the solvent is water, THF, methanol, ethanol, isopropanol, n-butanol, methyl ethyl ketone, or a combination thereof. In some embodiments, the solvent is water, ethanol, or a combination thereof. In some embodiments, R1 is p-methoxybenzyl. In some embodiments, the compound of formula (d) is (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid (Intermediate 6). In some embodiments, the compound of Formula (e) is ( R _a ) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) (Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitro Phenyl) ethylamine (1: 1 salt) (Intermediate 7): Intermediate 7

In some embodiments, the disclosed the synthesis of (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) -3 Method for-(3-methoxy-3-side oxypropyl) -1-methyl-1H-indole-2-formate (Intermediate 8): Intermediate 8 includes the steps of: (i) contacting a compound of formula (e) with an acid in the presence of a first solvent; (ii) separating a free acid component of a compound of formula (e); (iii) in The free acid is treated with a methylating agent or methanol in the second solvent; (iv) the protecting group is optionally removed in the third solvent; and (v) the ( R _a ) -methyl 6-chloro-7- (3 -(Hydroxymethyl) -1,5-dimethyl-1H-pyrazol-4-yl) -3- (3-methoxy-3-sideoxypropyl) -1-methyl-1H- Indole-2-formate (Intermediate 8). In some embodiments, the compound is having the formula R (e) of _a) -3- (2- carboxyethyl) -6-chloro-7- (3 - (((4-methoxybenzyl) oxy ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitrobenzene ) Ethylamine (1: 1 salt) (Intermediate 7). In some embodiments, the first solvent is tetrahydrofuran, toluene, or a combination thereof. In some embodiments, the methylating agent is DMF-DMA. In some embodiments, the second solvent is toluene. In some embodiments, the third solvent is methanol. In some embodiments, the protecting group is removed by acetyl chloride. In some embodiments, the protecting group is removed by acetamidine chloride in methanol. In some embodiments, the protecting group is removed by an acid, such as hydrochloric acid, which is formed in situ by the reaction of acetyl chloride and methanol.

In some embodiments, the disclosed the synthesis of (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) -3 -(3-hydroxypropyl) -1-methyl-1H-indole-2-formate (Intermediate 9): Intermediate 9 comprising the steps of: (i) making (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) 3- (3-methoxy-3- pendant propyl) -1-methyl-1H-indole-2-formate (intermediate 8) is contacted with a reducing agent in the presence of a solvent; and (Ii) Isolation of ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl-1H-pyrazol-4-yl) -3- (3-hydroxy (Propyl) -1-methyl-1H-indole-2-formate (Intermediate 9). In some embodiments, the reducing agent is diisobutylaluminum hydride. In some embodiments, the solvent is THF, hexane, or a combination thereof.

In some embodiments, a method for synthesizing methyl 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid (Intermediate 10) is disclosed: Intermediate 10 includes the following steps: (i) contacting dimethyl 1-methyl-1H-pyrazole-3,5-dicarboxylic acid with a reducing agent in the presence of a solvent; (ii) isolating 3- (hydroxymethyl) Methyl) -1-methyl-1H-pyrazole-5-carboxylic acid methyl ester; (iii) methyl 3- (hydroxymethyl) -1-methyl-1H-pyrazole-5-carboxylic acid with a chlorinating agent Contact; and (iv) isolated 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester (Intermediate 10). In some embodiments, the solvent is a mixture of methanol and 2-methyltetrahydrofuran. In some embodiments, the reducing agent is sodium borohydride or lithium borohydride. In some embodiments the reducing agent is sodium borohydride. In some embodiments, the solvent is methanol, ethanol, water, 2-methyltetrahydrofuran, dimethylacetamide, DCM, THF, cyclopentylmethyl ether, acetonitrile, or a mixture thereof. In some embodiments, the solvent is a mixture of methanol and 2-methyltetrahydrofuran.

In some embodiments, a method for synthesizing 3- (acetamidothio) naphthalene-1-yl acetate (Intermediate 11) is disclosed: Intermediate 11 includes the following steps: (i) contacting 4-hydroxynaphthalene-2-sulfonic acid with triphenylphosphine and iodine in a solvent; (ii) isolating 3-mercaptonaphthalene-1-ol; (iii) in Contacting 3-mercaptonaphthalene-1-ol with a hydrating agent in the presence of an amine base and a nucleophilic catalyst; and (iv) isolating 3- (acetamidothio) naphthalene-1-yl acetate (Intermediate 11) . In some embodiments, the solvent is acetonitrile. In some embodiments, the halogenating agent is acetic anhydride or acetamidine. In some embodiments, the saponifying agent is acetic anhydride. In some embodiments, the amine base is selected from triethylamine, pyridine, or diisopropylethylamine. In some embodiments, the amine base is triethylamine. In some embodiments, the nucleophilic catalyst is selected from the group consisting of 4-dimethylaminopyridine, pyridine, and N-methylimidazole. In some embodiments, the nucleophilic catalyst is 4-dimethylaminopyridine.

In some embodiments, the synthesis of methyl 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl-1H-pyrazole-3-carboxylic acid (Intermediate 12) is disclosed Method: Intermediate 12 includes the following steps: (i) 3- (acetamidothio) naphthalene-1-ylacetate (intermediate 11) and 5- (chloromethyl) -1- in the presence of a base and a solvent Methyl-1H-pyrazole-3-carboxylic acid methyl ester (Intermediate 10) in contact; and (ii) isolated 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl -1H-pyrazole-3-carboxylic acid ester (Intermediate 12). In some embodiments, the base is selected from potassium carbonate, lithium hydroxide, 1,8-diazabicyclo [5.4.0] undec-7-ene, sodium hydroxide, potassium hydroxide, and sodium ethoxide. In some embodiments, the base is potassium carbonate. In some embodiments, the solvent is selected from the group consisting of methanol, ethanol, water, and combinations thereof. In some embodiments, the solvent is methanol.

In some embodiments, the synthesis of 3-((((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol (intermediate 13) Method: Intermediate 13 includes the following steps: (i) methyl 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl-1H-pyrazole-3-carboxylic acid (intermediate 12) contact with a reducing agent in a solvent; and (ii) isolating 3-((((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene- 1-alcohol (Intermediate 13). In some embodiments, the reducing agent is diisobutylaluminum hydride. In some embodiments, the solvent is tetrahydrofuran, hexane, or a combination thereof.

In some embodiments, the synthesis of 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol (intermediate 14) Method: Intermediate 14 includes the following steps: (i) 3-(((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol ( Intermediate 13) in contact with a chlorinating agent; and (ii) isolating 3-((((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene- 1-alcohol (Intermediate 14). In some embodiments, the solvent is selected from the group consisting of THF, dichloromethane, dimethylformamide, and combinations thereof. In some embodiments, the solvent is dimethylformamide. In some embodiments, the chlorinating agent is methanesulfonyl chloride or thionyl chloride. In some embodiments, the chlorinating agent is methanesulfonyl chloride. In some embodiments, step (i) further comprises lithium chloride.

In some embodiments, the synthesis of 3-((((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate (Intermediate 15) Method: Intermediate 15 includes the following steps: (i) 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol ( Intermediate 14) is contacted with acetic anhydride, an optional amine base and a nucleophilic catalyst in a solvent; and (ii) isolates 3-(((3- (chloromethyl) -1-methyl-1H-pyrazole- 5-yl) methyl) thio) naphthalen-1-ylacetate (Intermediate 15). In some embodiments, the amine base is triethylamine. In some embodiments, the nucleophilic catalyst is selected from 4-dimethylaminopyridine, N-methylimidazole, or pyridine. In some embodiments, the nucleophilic catalyst is 4-dimethylaminopyridine. In some embodiments, the solvent is acetonitrile.

In some embodiments, the synthesis of 3-((((3-((acetamidothio) methyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1 -Method for acetic acid ester (Intermediate 16): Intermediate 16 includes the following steps: (i) 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylethyl Acid ester (Intermediate 15) in contact with potassium thioacetate in a solvent; and (ii) isolating 3-((((3-((acetamidinethio) methyl) -1-methyl-1H-pyrazole- 5-yl) methyl) thio) naphthalen-1-ylacetate (Intermediate 16). In some embodiments, the solvent is acetonitrile.

In some embodiments, the disclosed the synthesis of _{(R a) - (+)} - 17- methyl-28-chloro -5,13,14,22- four-oxa-2,9-dithia -5 , 6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37 ), 4 (38), 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formate Intermediate 17 comprising the steps of: (i) making (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) 3- (3-hydroxypropyl) -1-methyl-1H-indole-2-formate (Intermediate 9) is contacted with a sulfonating reagent in the presence of an aprotic base and a solvent to form a first Solution; (ii) adding an iodide salt to the first solution; (iii) isolating 6-chloro-7- [3- (iodomethyl) -1,5-dimethyl-pyrazol-4-yl]- 1-methyl-3- (3-methylsulfonyloxypropyl) indole-2-carboxylic acid methyl ester; (iv) make 3-((((3-((ethylamidothio) methyl)- 1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate (Intermediate 16) is contacted with a methanol solution of sodium methoxide to form a second solution; (v) the 6-chloro-7- [3- (iodomethyl) -1,5-dimethyl-pyrazol-4-yl] -1-methyl-3- (3-methylsulfonyloxypropyl) indole-2-carboxylate is added to the second solution; and (vi) separating _{(R a) - (+)} - 17- methyl-28-chloro-oxa four -5,13,14,22- -2,9-dithia-5,6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^{30, 35} ] 38 carbons-1 (37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formate . In some embodiments, the aprotic base is diisopropylethylamine or N-methylmorpholine. In some embodiments, the aprotic base is diisopropylethylamine. In some embodiments, the sulfonating agent is selected from the group consisting of methanesulfonyl anhydride, methanesulfonyl chloride, and p-toluenesulfonic anhydride. In some embodiments, the sulfonating agent is methanesulfonyl anhydride. In some embodiments, the solvent is an aprotic solvent. In some embodiments, the aprotic solvent is tetrahydrofuran. In some embodiments, the iodide salt is lithium iodide.

In some embodiments, a method for synthesizing Compound 1 is disclosed: Compound 1 includes the following steps: (i) making ( R _a )-(+)-methyl 17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia- 5,6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formate in contact with an ester hydrolysis reagent in a solvent, and ( ii) Separation ( R _a )-(+)-17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13,22 -Pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), 6 , 11,14,16,18,20,23,29,31,33,35-tridecene-23-carboxylic acid. In some embodiments, the ester hydrolysis reagent is a hydroxide base selected from the group consisting of lithium hydroxide, sodium hydroxide, and potassium hydroxide. In some embodiments, the base is sodium hydroxide. In some embodiments, the solvent is selected from the group consisting of methanol, ethanol, isopropanol, DMSO, water, and combinations thereof.

The term "isolated" refers to any suitable method for obtaining a desired compound from a reaction mixture. The term "isolated" includes extraction, filtration, drying, crystallization, evaporation, chromatography (eg, HPLC, column chromatography), metal removal, and the like. The term "isolated" includes methods for obtaining unpurified and purified compounds from a reaction mixture. In some embodiments, the compound can be isolated from the reaction mixture by extraction, and the next step in the reaction can be performed even if the compound has not been purified or removed from the solvent. Those skilled in the art will be able to determine a suitable method for isolating the desired compound from the reaction mixture.

The term "protecting group" includes hydroxyl protecting groups such as benzyl, p-methoxybenzyl (PMB), tetrahydropiperanyl (THP), p-methoxyphenyl (PMP), and tertiary butyl di Methylsilyl (TBDMS), , , , , , , , , and many more. Those skilled in the art will be able to identify suitable hydroxyl protecting groups without undue experimentation.

The term "protecting group precursor" includes reagents that add a protecting group to a desired moiety. In some embodiments, the protecting group precursor is a hydroxyl protecting group precursor, such as benzyl chloride, 4-methoxyphenol, bromomethylpyrazole, dihydropiperan, p-methoxyphenol, chloroform Triazole, tert-butyldimethylchlorosilane, tert-butyldimethylchlorosilane and 1- (chloromethyl) -4-methoxybenzene. Those skilled in the art will be able to easily identify suitable protecting group precursors without undue experimentation.

The term "reducing agent" includes lithium aluminum hydride, sodium borohydride, diisobutylaluminum hydride, and the like.

The term "solvent" includes non-polar solvents, aprotic solvents, and protic solvents. The term "non-polar solvent" includes pentane, cyclopentane, hexane, heptane, cyclohexane, benzene, toluene, chloroform, diethyl ether, cyclopentyl methyl ether, tertiary butyl methyl ether, tertiary pentyl methyl Ether, dichloromethane and methyl ethyl ketone. The term "aprotic solvent" includes tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, 2-methyltetrahydrofuran, 1,4-difluorene, dimethylacetamide, and dimethylmethylene. The term "protic solvent" includes n-butanol, isopropanol, n-propanol, ethanol, methanol, acetic acid, and water. In some embodiments, the solvent may include a combination of any of the foregoing solvents. Those skilled in the art will often be able to determine the appropriate solvent or combination of solvents for a particular reaction.

The term "brominating agent" includes 1,3-dibromo-5,5-dimethylhydantoin (DBDMH) and N-bromosuccinimide (NBS).

The term "diazotizing agent" includes sodium nitrite (NaNO ₂ ), calcium nitrite (Ca (NO ₂ ) ₂ ), and potassium nitrite (KNO ₂ ).

The term "metalizing agent" includes n-BuLi.

The term "ester hydrolyzing agent" includes sodium hydroxide (NaOH), potassium hydroxide (KOH), lithium hydroxide (LiOH), sodium chloride (NaCl), and lithium iodide (LiI).

The term "base" includes potassium acetate (KOAc), potassium carbonate (K ₂ CO ₃ ), sodium carbonate (Na ₂ CO ₃ ), potassium phosphate (K ₃ PO ₄ ), potassium hydroxide, sodium hydroxide, bis (trimethyl ether) Silyl) sodium sulfonamide (NaHMDS), bis (trimethylsilyl) sulfonamide (LiHMDS), sodium hydride (NaH), tributylammonium hydrogen sulfate, n-butyllithium, tributyllithium t-BuLi), magnesium, zinc, lithium hydroxide, lithium diisopropylamine (LDA), sodium amine (NaNH ₂ ), potassium tert-butoxide, pyridine, triethylamine (TEA), diisopropyl Ethylamine (DIEA), 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU), sodium methoxide (NaOMe), and sodium ethoxide (NaOEt). The term "base" also includes basic phase transfer catalysts, such as tetrabutylammonium hydrogen sulfate (Bu ₄ · HSO ₄ ), benzyltrimethylammonium chloride, polyethylene glycol and its derivatives, 18-crown ether- 6 and other crown ethers.

The term "acid" includes hydrochloric acid (HCl), sulfuric acid (H ₂ SO ₄ ), p-toluenesulfonic acid (p-TsOH), methanesulfonic acid, and acetic acid. In some embodiments, the acid is concentrated.

The term "alcohol" includes methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol and tertiary butanol.

The term "reducing agent" includes lithium aluminum hydride (LAH), borane-dimethylsulfide complex, borane-tetrahydrofuran complex, diisobutylaluminum hydride (DIBAL), lithium borohydride (LiBH ₄ ) , Sodium bis (2-methoxyethoxy) aluminum hydride (Red-Al®) and sodium borohydride (NaBH ₄ ).

The term "methylating agent" includes methyl iodide, dimethyl sulfate, and dimethylformamide-dimethylacetal (DMF-DMA).

The term "halogenating agent" includes acetic anhydride and acetamidine.

The term "nucleophilic catalyst" includes dimethylaminopyridine (DMAP), pyridine and N-methylimidazole.

The term "amine base" includes triethylamine, pyridine and diisopropylethylamine.

The term "sulfonating agent" includes toluenesulfonic anhydride, mesylate chloride, p-toluenesulfonyl chloride and mesylate anhydride.

The term "iodide salt" includes lithium iodide, sodium iodide, and potassium iodide.

In some embodiments, the term "palladium catalyst" includes 1,1'-bis (di-tertiary-butylphosphine) ferrocene palladium dichloride, [1,1'-bis (tertiary-butylphosphine) diamine Ferrocene] Palladium (II) dichloride (Pd 188, PdCl ₂ (dtbpf)), ( R )-(-)-4,12-bis (diphenylphosphine)-[2.2]- -Phanephos), ( S )-(-)-4,12-bis (diphenylphosphine)-[2.2] -p-cyclophane (S-Phanephos), (2-dicyclohexylphosphine-2 ′, 4 ′ , 6′-triisopropyl-1,1′-biphenyl) [2- (2′-amino-1,1′-biphenyl)] palladium (II) methanesulfonate (XPhos-G3-ring Palladium, XPhos-Pd-G3), (2-dicyclohexylphosphine-2 ′, 6′-diisopropoxy-1,1′-biphenyl) [2- (2′-amino-1,1 '-Biphenyl)] Palladium (II) mesylate (RuPhos-G3-cyclopalladium, RuPhos-Pd-G3), (2-dicyclohexylphosphine-2', 6'-diisopropoxy-1 , 1′-biphenyl) [2- (2′-amino-1,1′-biphenyl)] palladium (II) mesylate (RuPhos-G3-cyclopalladium, RuPhos-Pd-G3), [ (2-Dicyclohexylphosphine-3,6-dimethoxy-2 ′, 4 ′, 6′-triisopropyl-1,1′-biphenyl) -2- (2′-amino-1 , 1′-biphenyl)] palladium (II) mesylate (BrettPhos G3), tBuXPhos-Pd-G3, [(2-di-tertiary-butylphosphine-2 ′, 4 ′, 6′-trisiso Propyl-1,1 -Biphenyl) -2- (2′-amino-1,1′-biphenyl)] palladium (II) mesylate (tBu XPhos G3), tetrakis (triphenylphosphine) palladium (0) and bis (Di-tert-butyl (4-dimethylaminophenyl) phosphine) Palladium (II) dichloride

In some embodiments, Compound 1 can be synthesized as shown in Schemes I-VII: Scheme I A = i) NaNO ₂ , aqueous HCl; ii) 2-oxocyclopentane-1-carboxylic acid methyl ester, aqueous KOAc; iii) H ₂ SO ₄ , MeOH B = H ₂ SO ₄ , MeOH scheme II C = i) LiAlH ₄ , THF; ii) NBS iii) PMBCl, KOH, Bu ₄ NHSO ₄ , THF D = i) n-BuLi, THF; ii) i-PrOBPin scheme III E = Pd (di-t-BPF) Cl ₂ , K ₂ CO ₃ , 1,4-difluorene, H ₂ OF = i) DMF-DMA, PhMe; ii) Aqueous NaOH, MeOH G = (1R) -1 -(2-nitrophenyl) ethylamine hydrochloride, NaOH, EtOH, H ₂ O. H = i) aqueous HCl, PhMe, THF; ii) DMF-DMA, PhMe; iii) AcCl, MeOH I = iBu ₂ AlH, THF scheme IV J = i) NaBH ₄ , MeOH, MeTHF Scheme Ⅴ K = i) PPh ₃ , I ₂ , MeCN; ii) Ac2O, Et ₃ N, DMAP scheme Ⅵ L = K ₂ CO ₃ , MeOH M = i-Bu ₂ AlH, THF N = MeSO ₂ Cl, LiCl, DMF O = Ac ₂ O, DMAP, MeCN P = KSAc, MeCN Scheme VII Q = i) Intermediate 9, (MeSO ₂ ) ₂ O, i-Pr ₂ NEt; ii) LiI, MeCN; iii) In a separate container, intermediate 16, NaOMe, MeOH; iv) solution from iii) Added to compounds in MeCN from ii; v) Added to hot DMSO R = NaOH, DMSO, EtOH, H ₂ O [Example]

Aspects of this disclosure can be further defined by reference to the following non-limiting examples, which describe in detail the preparation of certain compounds and intermediates of this disclosure and methods for using the compounds of this disclosure. It should be clear to those skilled in the art that many modifications to both materials and methods can be practiced without departing from the scope of this disclosure.

Unless otherwise stated: (i) Unless otherwise stated, synthesis is performed at ambient temperature (in the range of 17ºC to 25ºC) and in an atmosphere of an inert gas such as nitrogen; (ii) by rotary evaporation or the use of Genevac equipment or The Biotage v10 evaporator is evaporated under reduced pressure; (iii) Kromasil® 60-10-SIL silica (10 μm particles, 10 μm particles, on an automated Novasep Hipersep® or Teledyne Isco CombiFlash® Rf or Teledyne Isco CombiFlash® Companion® system) 60Å pore size) or silica gel purification using pre-installed RediSep Rf Gold ™ silica columns (20-40 μm, spherical particles), GraceResolv ™ filters (Davisil® silica) or Silicycle filters (40-63 μm) . (Iv) Performed on a Waters X5 SFC-MS with UV detection or a Waters UPC2 SFC-MS with UV and ELSD detection or an Agilent 1100 HPLC system with UV detection Sex analysis chromatography. (V) If present, the yield is not necessarily the achievable maximum; (vi) Usually, the structure of the isolated compound is confirmed by NMR spectroscopy; using the solvent residual peak as an internal standard, the NMR chemical shift value is measured with a δ ruler [Using a Bruker Ultrashield Avance III 500MHz spectrometer equipped with a QCI cryoprobe, a Bruker Ultrashield Avance III 400 MHz spectrometer equipped with a BBFO probe, Bruker Avance 500 (500 MHz), Bruker Avance 400 (400 MHz), Bruker Avance 300 ( (300 MHz) or Bruker DRX (300 MHz) instrument for proton magnetic resonance spectroscopy]; unless otherwise stated, measurements were performed at 27ºC; the following abbreviations have been used: s, singlet; d, doublet; t, triplet; q, Quartet; m, multiplet; dd, double-doublet; ddd, double-doublet; bs, broad peak signal. (Vii) In general, Waters UPLC equipped with Waters SQ or QDa mass spectrometer is also used to characterize the separated compounds (column temperature 30ºC or 40ºC, UV = 220-300 nm or 210-400 nm or 190-400 nm, mass spectrum = ESI with positive / negative switching ESI), using a 97% A + 3% B to 3% A + 97% B solvent system, after 1.50 min (plus the total execution time of 1.70 min after equilibration back to the initial conditions, etc.) is performed at a flow rate of 1 mL / min, where A = 0.1% formic acid or 0.05% trifluoroacetic acid aqueous solution (for acidic operation) or 0.1% Aqueous ammonium hydroxide solution (for alkaline operation) and B = acetonitrile. For acid analysis, use Waters Acquity HSS T3 (1.8 μm, 2.1 × 50 mm or 2.1 × 30 mm) or Waters Acquity BEH C18 (1.7 μm, 2.1 × 50 mm or 2.1 × 30 mm), for neutral-pH analysis, Waters Acquity BEH C18 (1.7 μm, 2.1 × 50 mm) column was used, and for alkaline analysis, the column used was Waters ( Waters) Acquity BEH C18 (1.7 μm, 2.1 × 50 mm or 2.1 × 30 mm). Alternatively, a solvent gradient of 2% to 98% B over 1.5 min (plus a total execution time of equilibration back to the initial conditions of 2 min) is used, where A = 0.1% formic acid in water and B = 0.1% formic acid in Solution (for acidic operation) or A = 0.1% ammonium hydroxide aqueous solution and B = acetonitrile (for alkaline operation). Alternatively, after 3.6 min (plus a total cycle time of 5.1 min after equilibration back to the initial conditions, etc.), use 92% A + 5% B + 3% C to 7% A + 90% B + 3% C Or 90% A + 5% B + 5% D to 5% A + 90% B + 5% D solvent gradients where A = water, B = acetonitrile, C = 1% TFA in water and D = 250 mM acetic acid Aqueous ammonium solution; unless otherwise stated, the reported molecular ions correspond to [M + H] +; for molecules with multiple isotopic patterns (Br, Cl, etc.), the reported values are obtained using the highest intensities of. (Viii) In general, the wt% purity of a compound is determined by proton NMR under quantitative conditions (relative to a suitable internal reference standard (eg, 1,2,4,5-tetrachloro-3-nitrobenzene, Maleic acid or benzyl benzoate)). (Ix) large scale reactions in reactors equipped with heat transfer jackets and maintained with appropriate auxiliary equipment; and (x) the following abbreviations have been used: MeCN acetonitrile aq. Aqueous Conc. Concentrated DCM dichloromethane di-t-BPF 1,1'-bis (di-tertiary-butylphosphine) ferrocene DIBAH Diisobutylaluminum hydride DMAP 4-dimethylaminopyridine DMF N, N-dimethylformamide DMF-DMA N, N -Dimethylformamide dimethyl acetal DMSO dimethylarylene ee Mirror image isomer excess ES electrospray mode HPLC high performance liquid chromatography IPA isopropanol LAH lithium aluminum hydride MS mass spectrometry MTBE methyl tertiary Ether NBS N-bromosuccinimide NMR nuclear magnetic resonance PMB 4-methoxybenzyl TFA trifluoroacetic acid THF tetrahydrofuran UPLC ultra-high performance liquid chromatography wt% weight percentage intermediate 1 : (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydradino) adipate

A mixture of 2-bromo-3-chloroaniline (2.00 kg, 9.69 mol), hydrochloric acid (36 wt%, 4.85 L, 58.1 mol) and water (5 L) was stirred for 1 h. The resulting solution was cooled to 0ºC, and then a solution of NaNO ₂ (702 g, 10.2 mol) in water (2.4 L) was gradually added over 1 h at 0ºC-5ºC. After stirring for 1 h, methyl 2- pendant cyclopentane-1-carboxylate (1.38 kg, 9.69 mol) was gradually added at 0ºC-5ºC. Then gradually add a solution of KOAc (13.3 kg, 136 mol) in water (20 L). The resulting solution was allowed to react for another 45 min at 0ºC-5ºC. The solution was then extracted three times with DCM (12 L each). The combined organic extracts were washed with brine (10 L) and then charged into another reactor containing a solution of concentrated sulfuric acid (4.75 kg, 48.5 mol) in MeOH (3.1 kg). The resulting solution was allowed to react at 10ºC-20ºC for 3 h. The solution was concentrated to about 8 L, and then two cycles of adding MeOH (18 L per cycle) and distilling off the solvent under reduced pressure (18 L per cycle) were completed. The resulting slurry was cooled to 0ºC-10ºC, stirred for 1 h, and then filtered. The solid was washed with MeOH (2 × 2L) and then dried under reduced pressure in an oven to give (E / Z) -dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydrazinyl) Adipate, ( Intermediate 1 , 3.3 kg, 94 wt%, 82%); m / z (ES +), [M + H] ⁺ = 391. ¹ H NMR (500 MHz, chloroform- d , 27ºC) δ 1.98 (m, 2H), 2.41 (t, 2H), 2.59 (t, 2H), 3.66 (s, 3H), 3.87 (s, 3H), 7.05 (dd, 1H), 7.17 – 7.23 (m, 1H), 7.49 (dd, 1H), 12.48 (bs, 1H). Intermediate 2 : 7-bromo-6-chloro-3- (3-methoxy-3- pendant propyl) -1H-indole-2-carboxylic acid methyl ester

(E / Z) -Dimethyl 2- (2- (2-bromo-3-chlorophenyl) hydrazino) adipate ( Intermediate 1 , 3.3 kg, 93.7 wt%, 7.9 mol) in The solution in concentrated sulfuric acid (8.4 kg, 84 mol) and MeOH (26 L) was stirred at 80ºC for 72 h. The reaction mixture was cooled to 0ºC. The resulting solid was collected by filtration, washed with MeOH (2 L), and then dried in a vacuum oven at 40 ° C to give 2.5 kg of solid. This solid was combined with another 3 batches prepared in the same manner on the same scale to give a total of 11.9 kg of unpurified product from 13.6 kg (93.7 wt%, 32.5 mol) of starting material.

Half of the unpurified product was added to stirred MeOH (36 L). The mixture was heated to 65ºC, and the resulting solution was held at 65ºC for 1 h, and then cooled to 0ºC. The resulting slurry was stirred for 1 h at 0ºC and then filtered. The filter cake was washed with MeOH (3 L) and dried in a vacuum oven at 40 ° C. This process was repeated for the remaining materials and the materials were combined to give 9.7 kg of solids. 3.7 kg of it was mixed with activated carbon (0.74 kg), DCM (3.4 L) and MeOH (34 L), and the slurry was heated to 65ºC-70ºC for 1 h. The slurry was cooled to 55ºC and filtered. The filter cake was washed with DCM (10 L), and the combined filtrate was concentrated to about 8 L by distilling off the solvent under vacuum. Two cycles of adding MeOH (10 L) and distilling off the solvent (10 L) under vacuum were completed, and the resulting slurry was then combined with other similarly prepared slurry from the remaining solids. The combined slurry was cooled to 0ºC and stirred for 1 h, and then filtered. The filter cake was washed with MeOH (3 L) and dried in a 40 ° C vacuum oven to give 7-bromo-6-chloro-3- (3-methoxy-3- pendantoxypropyl) -1H-indole- Methyl 2-formate ( Intermediate 2 , 9.4 kg, 97.7 wt%, 76%); m / z (ES +), [M + H] ⁺ = 374. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC), δ 2.60 (t, 2H), 3.26 (t, 2H), 3.53 (s, 3H), 3.89 (s, 3H), 7.28 (d, 1H) , 7.73 (d, 1H), 11.62 (bs, 1H). Intermediate 3 : 4-bromo-3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole

At 4ºC-23ºC, LAH (1.05 M in THF, 15.0 kg, 17.4 mol) was gradually added to ethyl 1,5-dimethyl-1H-pyrazole-3-carboxylic acid (5.33 kg) over 1.5 h. 31.7 mol) in a stirred solution of THF (10.7 L), followed by the addition of THF (1.0 L). After 30 min, the solution was cooled to 15ºC. While continuing to cool, a solution of water (0.66 L, 37 mol) in THF (1.9 L) was gradually added over 20 min. Aqueous NaOH (15 wt%, 0.66 L, 2.8 mol) was then added over a few minutes, followed by water (2.0 L). The resulting slurry was stirred at 4ºC-11ºC for 20 minutes, and then filtered under suction. The collected solid was washed four times with THF (10.7 L per wash) to give (1,5-dimethyl-1H-pyrazol-3-yl) methanol as a solution in the collected filtrate.

The reactor was flushed with 1 M HCl, water and THF, after which the filtrate was refilled. Fill the solution with three parts of NBS (1.82 kg each, 99.4 wt%, total 30.6 mol), stir at 19ºC-27ºC between the filling parts for 7-8 minutes, and then stir at 21ºC-28ºC for 45 minutes . Then add a solution made of Na ₂ SO ₃ (0.81 kg, 99wt%, 6.4 mol), NaOH (50wt% in water, 4.6 kg, 57 mol) and water (16 L), and stir the resulting mixture at 25ºC-26ºC 10 min. The layers were separated and the lower layer was washed with THF (16 L). The upper layers were combined and evaporated to dryness to give (4-bromo-1,5-dimethyl-1H-pyrazol-3-yl) methanol as a solid.

This unpurified (4-bromo-1,5-dimethyl-1H-pyrazol-3-yl) methanol was re-dissolved in THF (18.8 L) and heated to 50 ° C with stirring, then tetrabutyl was added Ammonium bisulfate (0.32 kg, 0.95 mol), 1- (chloromethyl) -4-methoxybenzene (5.4 L, 97.6% by weight, 40 mol) and THF (2.8 L) line washing solution. At 47ºC-57ºC, gradually add KOH (45wt% in water, 13.7 L, 159 mol) to the vigorously stirred mixture over 45 min, and then continue stirring at 55ºC-50ºC for 4 h. The mixture was then cooled to 20ºC and kept under stirring for 63 h. Reheat the mixture to 50ºC, add water (18.7 L) and stir the mixture for 10 min. The lower layer was removed and the solution remaining in the reactor was cooled to 20ºC. 4-Bromo-3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole (0.01 kg, 0.03 mol) was added as a seed crystal and subsequently Heptane (32 L) was gradually added over 45 min, during which time crystallization began. The slurry was stirred at 20ºC for 30 min, then gradually filled with more heptane (22 L) over 45 min and cooled to 0ºC. After 17 h, more heptane (11 L) was added. After another 1 h at 0ºC, the slurry was filtered with suction. The filter cake was washed with a cooled (0ºC) mixture of heptane (17 L) and THF (4.3 L), and then dried in a 40ºC vacuum oven to give 4-bromo-3-(((4-methoxybenzyl Group) oxy) methyl) -1,5-dimethyl-1H-pyrazole ( Intermediate 3 , 8.78 kg, 96 wt%, 82%); m / z (ES +), [M + H] ⁺ = 325. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 2.22 (s, 3H), 3.74 (s, 3H), 3.75 (s, 3H), 4.32 (s, 2H), 4.39 (s, 2H), 6.87 – 6.93 (m, 2H), 7.22 – 7.27 (m, 2H). Intermediate 4 : 3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-4- (4,4,5,5-tetramethyl-1,3 , 2-dioxolane-2-yl) -1H-pyrazole

4-Bromo-3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole ( intermediate at -73ºC to -66ºC over 1.5 h 3 , 5.27 kg, 96 wt%, 15.6 mol) in a stirred slurry of THF (43 L) was gradually added butyllithium (15 wt% in hexane, 7.27 kg, 17.3 mol). The resulting solution was stirred at -77ºC to -66ºC for 1.7 h, and then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxolane was added over 15 min. (3.4 L, 16 mol), followed by THF (3.0 L) line wash. Stir the solution for 1.5 h at -77ºC to -63ºC, then gradually add a solution of acetic acid (0.89 L, 16 mol) in toluene (25.3 L) at -77ºC to -58ºC over 15 min. The mixture was then warmed to 20ºC, then heated and the solvent (48 L) was distilled off at 67ºC-82ºC (atmospheric pressure). The mixture was cooled to 65ºC, water (25.5 L) was added and the mixture was stirred for 10 min. Remove the lower layer and distill off more solvent (25.4 L) at 72ºC-119ºC (atmospheric pressure; final vapor temperature of 108ºC). The resulting solution was cooled to 40ºC and diluted with heptane (50.6 L) over 10 min, during which time the mixture was cooled to 21ºC and spontaneous crystallization began. While 3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-4- (4,4,5,5-tetramethyl-1,3,2 -Dioxolane-2-yl) -1H-pyrazole (29 g) was added as a seed crystal. The slurry was stirred at about 21ºC for 0.6 h, cooled to about -5ºC over 1.5 h, and then kept at this temperature overnight (18 h). The slurry was filtered by suction, and the filter cake was washed with cold (about 0ºC) heptane, and then dried in a 40ºC vacuum oven to give 3-(((4-methoxybenzyl) oxy) methyl) -1 , 5-dimethyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxolane-2-yl) -1H-pyrazole ( intermediate 4 , 4.50 kg, 99 wt%, 77%); m / z (ES +), [M + H] ⁺ = 373. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 1.23 (s, 12H), 2.34 (s, 3H), 3.68 (s, 3H), 3.73 (s, 3H), 4.40 (s, 2H), 4.42 (s, 2H), 6.85 – 6.91 (m, 2H), 7.20 – 7.25 (m, 2H). Intermediate 5 : (±) -methyl 6-chloro-3- (3-methoxy-3- pendant propyl) -7- (3-(((4-methoxybenzyl) oxy ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1H-indole-2-formate

A 100 L reactor was charged with 3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-4- (4,4,5,5-tetra Methyl-1,3,2-dioxolane-2-yl) -1H-pyrazole ( intermediate 4 , 3.30 kg, 99 wt%, 8.78 mol), 7-bromo-6-chloro- 3- (3-methoxy-3- pendantoxypropyl) -1H-indole-2-carboxylic acid methyl ester ( intermediate 2 , 3.20 kg, 97.7 wt%, 8.35 mol), K ₂ CO ₃ (1.60 kg, 11.6 mol) and 1,1'-bis (tertiary-butylphosphino) ferrocene palladium dichloride (0.132 kg, 0.203 mol). The reactor headspace was then evacuated and refilled with nitrogen three times. 1,4-Diamidine (26.3 L) and water (3.3 L) were bubbled with nitrogen under reduced pressure for 5-10 min, and then they were added sequentially, and the resulting slurry was heated at 80 ° C for 5 h with stirring. The reaction mixture was cooled to 20ºC and held overnight (16 h) before being diluted with MTBE (33 L) and water (33 L). Add N -acetamidine cysteine (0.165 kg, 1.01 mol) and stir the mixture for 15 min. The lower layer is removed once it has settled. The upper layer was then washed with hydrochloric acid (37 wt%, 2.7 L, 33 mol) in water (30 L) and then with water (32 L) to give (±) -methyl 6-chloro-3- (3-formaldehyde) Oxy-3- pendant oxypropyl) -7- (3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole-4- Solution) -1H-indole-2-formate ( Intermediate 5 , 22.9 kg, 19.1 wt%, 97%) in MTBE. Intermediate 6 : (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) methyl) -1,5-di (Methyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid

(±) -methyl 6-chloro-3- (3-methoxy-3- pendantoxypropyl) -7- (3-(((4-methoxybenzyl) oxy) methyl ) -1,5-Dimethyl-1H-pyrazol-4-yl) -1H-indole-2-formate ( Intermediate 5 , 22.94 kg, 19.1 wt%, 8.12 mol) in MTBE solution at 20ºC It was stirred with mercaptopropyl-functionalized palladium dioxide (Quadrasil ^™ MP, 1.1 kg) and toluene (44 L) for 1 h. The slurry was filtered and the solid waste was washed with toluene (13 L). The combined filtrates were concentrated by removing the solvent (44 L) at 75ºC-107ºC. Then add DMF-DMA (5.5 L, 41 mol) at 91ºC. The solution was heated to reflux for 22 h at approximately 108ºC, during which four solvents (2, 4.5, 4.5, and 4.4 L) were distilled off (removed immediately after reaching reflux, and then 2.0, 3.4 and 4.8 h distilled off), and then cooled to 55ºC. Fill a solution of NaCl (2.2 kg) in water (20 L), then remove the resulting two-phase mixture from the container and filter it back through a 5 μm line filter. The sieved mixture was stirred at 50ºC-53ºC for 10 min, and once settled, the lower layer was removed. Then MeOH (22 L) and aqueous NaOH (3.3 kg, 50 wt%, 41 mol) in water (18 L) were added, and the resulting two-phase mixture was stirred at 55ºC for 3 h. After settling, the layers were removed to separate the containers. The lower layer (containing product) is then returned to the reactor and stirred at 55ºC. Then gradually add a solution of acetic acid (4.6 kg, 77 mol) in water (4.4 L) over 25 min, followed by 3- (2-carboxyethyl) -6-chloro-7- (3-(((4- (Methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid (0.02 kg, 0.04 mol ) Crystal seed. The mixture was held at 49ºC-56ºC for 2 h, cooled to 20ºC over 2 h, and then maintained at 20ºC for 13 h. The resulting slurry was filtered by suction. Rinse the reactor and filter cake with water (11 L), then partially dry on the filter by continuing to draw at ambient temperature for 6 h, and completely dry in a 40ºC vacuum oven to give (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazole-4- Group) -1-methyl-1H-indole-2-carboxylic acid ( Intermediate 6 , 3.78 kg, 99.0 wt%, 86%); m / z (ES +), [M + H] ⁺ = 526. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 2.01 (s, 3H), 2.50 – 2.57 (m, 2H), 3.18 – 3.29 (m, 2H), 3.40 (s, 3H), 3.68 (s , 3H), 3.82 (s, 3H), 4.08 (d, 1H), 4.14 (d, 1H), 4.19 (d, 1H), 4.21 (d, 1H), 6.64 – 6.69 (m, 2H), 6.69- 6.74 (m, 2H), 7.25 (d, 1H), 7.75 (d, 1H), 12.71 (bs, 1H). Intermediate _{7: (R a) -3- (} 2- carboxyethyl) -6-chloro-7- (3 - (((4-methoxybenzyl) oxy) methyl) -1,5 Dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitrophenyl) ethylamine (1: 1 salt )

Add NaOH (50 wt% in water, 0.95 L, 18 mol) to (±) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxy Benzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid ( intermediate 6 , 9.38 kg, 97.5 wt%, 17.4 mol) and (1 R ) -1- (2-nitrophenyl) ethylamine hydrochloride (2.48 kg, 91 wt%, 11.1 mol) in water (7.4 kg) and ethanol (64 L ), Followed by rinsing with ethanol (1.8 L). When the resulting solution is heated to 78ºC, spontaneous crystallization of the product begins (at or below 41ºC). After heating to 78ºC, the three cycles of cooling the slurry to 63ºC in 1.3 h, heating back to 78ºC in 0.6 h, and maintaining for 10 min were completed. The slurry was then cooled to 63ºC over 1.2 h, then to 20ºC over 1.6 h, then maintained at 20ºC overnight, and then filtered with suction. Rinse the reactor and filter cake with ethanol (18 L). Once fully deliquored, the filter cake was returned to the reactor. (The solid usually has about 92% ee of the desired diacid). Fill with water (7.3 L) and ethanol (66 L) and heat the reactor contents to 78 ° C with stirring. The resulting slurry was cooled to 63ºC over 1.3 h, then cooled to 20ºC over 1.6 h, then kept at 20ºC overnight, and then filtered with suction. Rinse the reactor and filter cake with ethanol (18 L). The filter cake was dried in a 40ºC vacuum oven to give ( R _a ) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxybenzyl) oxy) ) Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitrobenzene Ethyl) Ethylamine (1: 1 salt) ( Intermediate 7 , 5.23 kg, 97.5 wt%, 99.0% ee, 7.36 mol) Yield is 42.3%; m / z (ES +), [M + H] ⁺ = 526 (Acid), 167 (amine). ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 1.49 (d, 3H), 2.00 (s, 3H), 2.61 (t, 2H), 3.10 – 3.22 (m, 2H), 3.38 (s, 3H ), 3.68 (s, 3H), 4.11 (d, 1H), 4.14 (d, 1H), 4.17 (d, 1H), 4.22 (d, 1H), 4.66 (q, 1H), 6.67 – 6.72 (m, 2H), 6.75 – 6.80 (m, 2H), 7.18 (d, 1H), 7.54 – 7.60 (m, 1H), 7.63 (d, 1H), 7.76 – 7.82 (m, 1H), 7.90 – 7.94 (m, 1H), 7.93 – 7.97 (m, 1H), 9.16 (bs, 3H). Chiral purity analysis HPLC method details: column = Chiralpak AD-H (4.6 × 250 mm, 5 μm); temperature = 25ºC; mobile phase = 70: 30 hexane: 0.2% TFA in ethanol (by volume), flow rate 1.0 mL / min; UV at 254 nm by detection; injection volume = 10 μL (which can be adjusted to achieve proper detection limit); retention time R _a and S _a enantiomers were 4.8 and 13.7 min. Intermediate 8 : ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl-1H-pyrazol-4-yl) -3- (3-methyl (Oxy-3- pendant oxypropyl) -1-methyl-1H-indole-2-formate

Add hydrochloric acid (37 wt%, 0.77 L, 9.2 mol) to ( R _a ) -3- (2-carboxyethyl) -6-chloro-7- (3-(((4-methoxy Benzyl) oxy) methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -1-methyl-1H-indole-2-carboxylic acid- (1 R ) -1- (2-nitrophenyl) ethylamine (1: 1 salt) ( Intermediate 7 , 5.23 kg, 97.5 wt%, 7.36 mol) in a stirred slurry of THF (20.5 L) and water (20.5 L). After 5 min, toluene (41 L) was added and the mixture was stirred for 10 min. Remove the lower layer, dilute the upper layer with toluene (20.5 L), and concentrate by removing the solvent (60 L) under reduced pressure (590 mbar) at 48ºC-92ºC. DMF-DMA (3.45 L, 25.8 mol) was added to the resulting mixture at 90ºC to obtain a solution. The solution was heated to reflux, maintained at reflux (98ºC) for 8 h, then cooled to 49ºC and held for 16 h. Add DMF-DMA (1.0 L, 7.5 mol) and reflux the mixture at 100ºC for another 3.6 h before cooling to 50ºC. Water (12.7 L) was added and the mixture was stirred for 15 min. Remove the lower layer. MeOH (20 L) was added to the upper layer, and then acetamidine (2.15 L, 29.9 mol) was gradually added to the stirred solution for 10 min. The solution was heated at 60ºC for 21 h and then transferred to a container. Fill the empty reactor with toluene (10 L) and K ₂ CO ₃ (2.56 kg, 18.4 mol) in water (20.5 L), and heat the mixture to 55ºC. The completed reaction solution was then gradually returned to the reactor over 20 min, followed by rinsing with toluene (10 L). Stir at 55ºC for another 10 minutes and stop stirring. Once settled, remove the lower layer. The upper layer was concentrated by removing the solvent (21 L) under reduced pressure (540 mbar) at 55ºC-93ºC. Dilute the solution with heptane (10 L) at 50ºC and inoculate ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl-1H-pyridine Azol-4-yl) -3- (3-methoxy-3- pendantoxypropyl) -1-methyl-1H-indole-2-formate (16 g, 0.037 mol). After 1 h of crystallization was established at 50ºC, more heptane (20 L) was gradually added over 1 h. The slurry was then cooled to 20ºC over 2 h and stirred for 65 h, and then filtered with suction. The filter cake was washed with heptane (10 L) and then dried in a 40 ° C vacuum oven to give ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-di Methyl-1H-pyrazol-4-yl) -3- (3-methoxy-3- pendantoxypropyl) -1-methyl-1H-indole-2-formate ( Intermediate 8 , 2.85 kg, 98 wt%, 87%); m / z (ES +), [M + H] ⁺ = 434. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 1.95 (s, 3H), 2.57 – 2.64 (m, 2H), 3.17 – 3.33 (m, 2H), 3.44 (s, 3H), 3.57 (s , 3H), 3.79 (s, 3H), 3.84 (s, 3H), 4.13 (dd, 1H), 4.23 (dd, 1H), 4.72 (dd, 1H), 7.26 (d, 1H), 7.72 (d, 1H). Intermediate 9: (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) -3- (3-hydroxy (Propyl) -1-methyl-1H-indole-2-formate

( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl-1H-pyrazol-4-yl) -3- (3-methoxy- 3-Phenoxypropyl) -1-methyl-1H-indole-2-formate ( Intermediate 8 , 2.25 kg, 98 wt%, 5.08 mol) was dissolved in THF (13.3 L), and The resulting solution was cooled to -45ºC. Then DIBAH (20.3wt% in hexane; 11.0 kg, 15.7 mol) was added at -41ºC or below -41ºC over 1.1 h. The reaction mixture was held at approximately -45ºC for an additional 4.4 h, during which additional three portions of DIBAH (20.3 wt% in hexane; 1.32, 0.25, and 0.07 kg; 1.88) were added after 1.4 h, 2.8 h, and 3.7 h , 0.36, and 0.10 mol), then IPA (2.2 L, 29 mol) was added to the reaction mixture, and then heated to 20ºC over 2 h and maintained at this temperature for 2.5 h.

Meanwhile, in another reactor, sodium potassium tartrate tetrahydrate (6.47 kg, 22.9 mol) and water (22 L) were charged. After stirring at 20ºC for several minutes, a solution was formed and then isopropyl acetate (22 L) was added. The resulting two-phase mixture was heated to 50ºC.

The ester reduction reaction mixture was transferred to a vigorously stirred mixture of aqueous tartrate and isopropyl acetate at 50ºC for 20 min, and then rinsed with THF (1.5 L). Continue vigorous stirring at 50ºC for 1.9 h. Remove the lower layer. The upper layer was washed with water (4.45 L), then removed and filtered back to the reactor through a 5 μm line filter, followed by line washing with isopropyl acetate (1.1 L). Concentrate the solution by distilling off the solvent (32 L) at 58ºC-74ºC (atmospheric pressure) and then cool to 20ºC. The solution was transferred to a smaller container, followed by line washing with isopropyl acetate (1.1 L), and then further concentrated (to 15 liters) by distillation to remove more solvent (15 L) at 73ºC-85ºC (atmospheric pressure). Approx. 9 L). The stirred solution was then cooled to 70ºC and inoculated with ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl-1H-pyrazol-4-yl)- 3- (3-hydroxypropyl) -1-methyl-1H-indole-2-formate (2 g, 97.9 wt%, 5 mmol), cooled to 20ºC over 1 hour and maintained at 20ºC for 24 h . The resulting slurry was filtered by suction. The filter cake was washed with isopropyl acetate (2.2 L) and then dried in a 40ºC vacuum oven to give ( R _a ) -methyl 6-chloro-7- (3- (hydroxymethyl) -1,5 -Dimethyl-1H-pyrazol-4-yl) -3- (3-hydroxypropyl) -1-methyl-1H-indole-2-formate ( Intermediate 9 , 1.76 kg, 98.0 wt %, 84%); m / z (ES +), [M + H] ⁺ = 406. ¹ H NMR (500 MHz, DMSO- d ₆ , 27 ºC) δ 1.68 – 1.77 (m, 2H), 1.96 (s, 3H), 2.93 – 3.05 (m, 2H), 3.43 (s, 3H), 3.42 – 3.48 (m, 2H), 3.80 (s, 3H), 3.84 (s, 3H), 4.14 (dd, 1H), 4.23 (dd, 1H), 4.48 (t, 1H), 4.71 (dd, 1H), 7.25 (d, 1H), 7.71 (d, 1H). Intermediate 10 : methyl 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid

Add sodium carbonate (2.14 kg, 20.2 mol) to stirred 1-methyl-1H-pyrazole-3,5-dicarboxylic acid (7.95 kg, 40.1 mol) in MeOH (80 L at 20ºC ± 5ºC) ) In solution. The slurry was stirred for 0.5 h and then filtered. The filter cake was washed with 2-methylTHF (16 L), and the filtrate was returned to the reactor, followed by the addition of 2-methylTHF (24 L). The solution was cooled to 15ºC ± 3ºC, and sodium borohydride (3.05 kg, 80.7 mol) was added in ten portions at 15ºC ± 3ºC. After adding NaBH ₄ , the mixture was stirred at 18ºC ± 3ºC for 2 h. Then quench by gradually filling acetone (16.4 kg, 283 mol) at 18ºC ± 3ºC, and then stir at 20ºC ± 5ºC for 1 h. Then slowly add aqueous HCl (37 wt%, about 8.6 kg, 87 mol), keep the temperature below 30ºC to adjust the pH to 2-3, and then stir the mixture for 1 h. Then saturated aqueous Na ₂ CO ₃ (about 4 L) was slowly added to adjust the pH to 5-6, and the mixture was stirred for 3 h. The mixture was filtered and the filter cake was washed with DCM (16 L). The filtrate was concentrated to about 20 L by distillation under reduced pressure, heated to no more than 40ºC, then diluted with DCM (40 L) and concentrated again by removing the solvent (about 40 L) under reduced pressure. Fill with DCM (80 μL) and pure water (32 μL) and stir the resulting mixture for at least 10 min. The lower (organic) phase was collected and the upper (aqueous) phase was extracted four times with more DCM (40 L / part). The combined organic phases were concentrated to about 20 L by vacuum distillation below 40 ° C. DCM (80 μL) was added and the solvent was removed by distillation (approximately 40 μL) to concentrate the solution again. The resulting 5- (hydroxymethyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester solution was then diluted with DCM (80 L) and cooled to 10ºC ± 5ºC, so while keeping the temperature below 15ºC Gradually add thionyl chloride (4.80 kg, 40.4 mol). The mixture was then stirred at 20ºC ± 5ºC for 1 h. The mixture was concentrated to no more than 20 L by vacuum distillation below 40ºC, then DCM (64 L) and pure water (80 L) were added. After phase separation, the lower (organic) phase was washed twice with aqueous Na ₂ CO ₃ (9 wt%, 80 L each), and then washed with pure water (80 L). The washed organic phase was concentrated to about 14 L by vacuum distillation below 40 ° C. Two cycles of slow addition of heptane (40 L) and vacuum distillation below 45 ° C to about 24 L are then completed. Slowly add more heptane (40 L) and stir the resulting slurry at 20ºC ± 5ºC for at least 0.5 h before filtering. The filter cake was washed with heptane (8.0 L) and then dried in a 45 ° C vacuum oven to give 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester ( Intermediate 10 , 4.87 kg, 64%); m / z (ES +), [M + H] ⁺ = 189. ¹ H NMR (500 MHz, CDCl ₃ , 27ºC) δ 3.91 (s, 3H), 3.99 (s, 3H), 4.59 (s, 2H), 6.82 (s, 1H). Intermediate 11 : 3- (Ethylthio) naphthalen-1-ylacetate

Mix a stirred mixture of MeCN (102 μL), sodium 4-hydroxynaphthalene-2-sulfonate (17.00 kg, 69.05 mol), triphenylphosphine (65.1 kg, 248 mol), and iodine (14.0 kg, 55.2 mol) at 80ºC. Heat at ± 5ºC for 6 h. Cool the mixture and stir at 0ºC ± 5ºC for at least 2 h. The waste solid was filtered off to give crude 3-mercaptonaphthalene-1-ol dissolved in the filtrate (also containing a large amount of triphenylphosphine oxide). The filtrate was refilled into the reactor, followed by DMAP (0.84 kg, 6.9 mol). Keep the temperature below 25ºC, gradually add triethylamine (21.0 kg, 207 mol), and then add acetic anhydride (17.6 kg, 173 mol). Stir the mixture at 15ºC-20ºC for 2 h, and then distill to less than 85 L under reduced pressure. Do not heat above 45ºC. Add DCM (85 L), and then distill the mixture under reduced pressure to less than 85 L. Do not heat above 40ºC. More DCM (170 L) was added, and the mixture was washed with water (170 L) and then with aqueous NaCl (17 wt%, 170 L). Distill the lower organic phase to less than 51 L under reduced pressure. Do not heat above 40ºC. Three cycles of diluting the solution with MeOH (85 L) and distilling less than 51 L under 40 ° C under reduced pressure were then completed to give crude 3- (acetamidinethio) naphthalene-1- in a yield of 71% Methyl acetate ( Intermediate 11 , 40.9 kg, 31.3 wt%, 49.2 mol) in methanol. A solution also containing a large amount of triphenylphosphine oxide was used directly for the preparation of intermediate 12 . Intermediate 12 : 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester

Stir 3- (acetamidothio) naphthalene-1-ylacetate ( intermediate 11 , 40.9 kg, 31.3 wt% in methanol, 49.2 mol), methanol (64 L), and K ₂ CO at 15ºC-20ºC ₃ (13.6 kg, 98.4 mol). Add 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester ( Intermediate 10 , 7.40 kg, 39.2 mol) in portions while keeping the temperature below 25ºC. The slurry was then stirred at 20ºC-25ºC for 2 h. In response to HPLC analysis, four additional portions of 5- (chloromethyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester ( Intermediate 10 , 0.46 kg, 2.4 mol / portion) were charged, and each portion was charged Stir for 1 h at 20ºC-25ºC. Add pure water (109 L) gradually at 15ºC-20ºC, stir the resulting mixture for at least 2 h, then let it stand for at least 3 h. The liquid was removed by a pipe, the settled viscous solid was left in the reactor, and then ethanol (25.5 L) was added thereto. Stir the mixture for at least 1 h at 15ºC ± 5ºC, and then filter the resulting slurry. The filter cake was washed with ethanol (6.4 L) and then dried in a 45ºC ± 5ºC vacuum oven to give 5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl- 1H-pyrazole-3-carboxylic acid methyl ester ( Intermediate 12 , 13.05 kg, 99.3 wt%, 81%); m / z (ES +), [M + H] ⁺ = 329. ¹ H NMR (500 MHz, DMSO, 27ºC) δ 3.72 (s, 3H), 3.90 (s, 3H), 4.39 (s, 2H), 6.63 (s, 1H), 6.79 (d, 1H), 7.28 – 7.32 (m, 1H), 7.38 (ddd, 1H), 7.45 (ddd, 1H), 7.72 (d, 1H), 8.07 (d, 1H), 10.55 (bs, 1H). Intermediate 13 : 3-(((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol

5-(((4-hydroxynaphthalen-2-yl) thio) methyl) -1-methyl-1H-pyrazole-3-carboxylic acid methyl ester ( Intermediate 12 , 12.7 kg, 99.3 at 30ºC ± 5ºC wt%, 38.4 mol) was dissolved in THF (254 L), and the solution was cooled to 15ºC ± 5ºC. Add DIBAH (1 M in hexane, 155 L, 155 mol) slowly while maintaining the temperature at 15ºC ± 5ºC. The mixture was then stirred at 20ºC ± 5ºC for 0.5 h, analyzed by HPLC, and then gradually transferred to aqueous hydrochloric acid (4 M, 114 L, 456 mol) at 5ºC-20ºC. Distill the two-phase mixture under vacuum below 40 ° C to no more than about 120 L. The resulting slurry was cooled to 15ºC ± 5ºC and then filtered. The filter cake was washed with pure water (25 L) and then refilled into the reactor with DCM (57 L) and THF (6.4 L). After stirring the mixture for at least 10 h at 20ºC ± 5ºC, filter it. The filter cake was washed with DCM (25 L) and then dried in a vacuum oven at 45ºC ± 5ºC to give 3-(((3- (hydroxymethyl) -1-methyl-1H-pyrazole-5- (Methyl) methyl) thio) naphthalene-1-ol ( Intermediate 13 , 10.45 kg, 95.5 wt%, 87%); m / z (ES +), [M + H] ⁺ = 301. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 3.77 (s, 3H), 4.28 (s, 2H), 4.37 (s, 2H), 4.87 (bs, 1H), 6.14 (s, 1H), 6.81 (d, 1H), 7.35 (d, 1H), 7.39 (ddd, 1H), 7.47 (ddd, 1H), 7.72 – 7.75 (m, 1H), 8.03 – 8.06 (m, 1H), 10.34 (s, 1H). Intermediate 14 : 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol

Methanesulfonyl chloride (6.28 kg, 54.8 mol) was gradually added to 3-(((3- (hydroxymethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene- In a stirred mixture of 1-alcohol ( Intermediate 13 , 10.30 kg, 95.5 wt%, 32.7 mol), anhydrous lithium chloride (2.91 kg, 68.6 mol), and DMF (51.5 L), while maintaining the temperature below 10ºC. Stir the mixture at 15ºC-20ºC for 2 h. Then EtOAc (155 L) was added, then pure water (155 L) was added and the mixture was mixed well. The lower layer was removed and the upper layer was washed twice with aqueous NaCl (17 wt%; 155 L each). The upper layer was then vacuum distilled to less than 50 L at less than 35ºC. Then slowly add heptane (155 L) at 30ºC ± 5ºC, and then cool the mixture to 0ºC-5ºC. After stirring the slurry for at least 1 h, it was filtered. The filter cake was washed with heptane (10.3 L) and then dried in a vacuum oven at 30ºC-35ºC to give 3-(((3- (chloromethyl) -1-methyl-1H-pyrazole-5 -Yl) methyl) thio) naphthalene-1-ol ( Intermediate 14 , 9.72 kg, 95.3 wt%, 89%); m / z (ES +), [M + H] ⁺ = 319. ¹ H NMR (500 MHz, CDCl ₃ , 27 ºC) δ 3.80 (s, 3H), 4.07 (s, 2H), 4.50 (s, 2H), 6.11 (s, 1H), 6.70 (d, 1H), 7.38 – 7.42 (m, 1H), 7.45 – 7.52 (m, 2H), 7.52 (bs, 1H), 7.68 – 7.74 (m, 1H), 8.16 – 8.20 (m, 1H). Intermediate 15 : 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate

Fill acetic anhydride (3.65 kg, 35.8 mol) into 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1 -A stirred mixture of alcohol ( Intermediate 14 , 9.51 kg, 95.3 wt%, 28.4 mol), DMAP (360 g, 2.95 mol) and MeCN (95 L), while maintaining the temperature below 25ºC. Stir the mixture at 15ºC-20ºC for 2 h. Then EtOAc (95 L) was added, followed by aqueous NaCl (10 wt%, 95 L). After thorough mixing, the lower layer was removed. The upper layer was washed with two additional portions of aqueous NaCl (10 wt%, 95 L each) and then subjected to three cycles of vacuum distillation at less than 40ºC to less than 29 L and then adding MTBE (95 L). Distill the mixture under vacuum below 40ºC to less than 48 L, stir at about 20ºC for at least 1 h, then cool to 0ºC-5ºC. Slowly add heptane (95 L), stir the slurry at 0ºC-5ºC for at least 1 h, and then filter. The filter cake was washed with heptane (17 L) and then dried in a vacuum oven at 35ºC-40ºC to give 3-(((3- (chloromethyl) -1-methyl-1H-pyrazole-5 -Yl) methyl) thio) naphthalene-1-ylacetate ( Intermediate 15 , 8.67 kg, 96.1 wt%, 81%); m / z (ES +), [M + H] ⁺ = 361. ¹ H NMR (500 MHz, CDCl ₃ , 27 ºC) δ 2.46 (s, 3H), 3.82 (s, 3H), 4.11 (s, 2H), 4.49 (s, 2H), 6.10 (s, 1H), 7.19 (d, 1H), 7.48 – 7.55 (m, 2H), 7.65 – 7.67 (m, 1H), 7.73 – 7.79 (m, 1H), 7.79 – 7.85 (m, 1H). Intermediate 16 : 3-(((3-((ethylamidothio) methyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate

Add potassium thioacetate (4.15 kg, 36.3 mol) to 3-(((3- (chloromethyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1 -Glycolate ( Intermediate 15, 8.62 kg, 96.1 wt%, 23.0 mol) and MeCN (86 L) in a mixture while maintaining the temperature below 25ºC. Stir the mixture at 15ºC-20ºC for 3 h. Then EtOAc (86 L) was added, followed by water (86 L). After thorough mixing, the lower layer was removed. The upper layer was washed with two portions of aqueous NaCl (15 wt%, 86 L each), then subjected to vacuum distillation at less than 40ºC to less than 29 L, then MTBE (86 L × 3, and 60 L in the final cycle) ) Of the four cycles. Stir the mixture at 30ºC-35ºC for at least 1 h, cool to below 0ºC, then stir at below 10ºC for at least 1 h, then slowly add heptane (86 L). Before filtering, cool the slurry to 0ºC-5ºC and keep it for at least 1 h. The filter cake was washed with heptane (17 L) and then dried in a vacuum oven at 35ºC-40ºC to give 3-(((3-((ethylamidothio) methyl) -1-methyl-1H -Pyrazol-5-yl) methyl) thio) naphthalene-1-yl acetate ( Intermediate 16 , 8.44 kg, 97 wt%, 89%); m / z (ES +), [M + H] ⁺ = 401. ¹ H NMR (500 MHz, DMSO- d ₆ , 27ºC) δ 2.28 (s, 3H), 2.45 (s, 3H), 3.75 (s, 3H), 3.91 (s, 2H), 4.42 (s, 2H), 6.04 (s, 1H), 7.36 (d, 1H), 7.53 (ddd, 1H), 7.58 (ddd, 1H), 7.82 (d, 1H), 7.85 (d, 1H), 7.90 (d, 1H). Intermediate 17 : ( R _a )-(+)-methyl 17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12, 13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38 ), 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formate

Add a solution of methanesulfonic anhydride (1.20 kg, 6.89 mol) in MeCN (2.5 L) to ( R _a ) -methyl 6-chloro-7- (3- (hydroxyl) at 0ºC-30ºC for 15 min. (Methyl) -1,5-dimethyl-1H-pyrazol-4-yl) -3- (3-hydroxypropyl) -1-methyl-1H-indole-2-formate ( intermediate 9 , 1.19 kg, 97.6 wt%, 2.86 mol) and a solution of N , N -diisopropylethylamine (1.5 L, 8.6 mol) in THF (8.7 L), followed by pipeline with MeCN (0.3 L) washing. The resulting solution was stirred at 20ºC for 5 h, and then cooled to about -5ºC. At the same time, LiI (1.53 kg, 11.4 mol) solution was prepared by adding (0.25 kg per portion) to MeCN (13 L) in portions below 30ºC. At approximately -5ºC, the LiI solution was gradually added to the mesylate reaction mixture over 20 min, followed by line washing with MeCN (1.2 L). The resulting slurry was warmed to 5ºC and stirred at this temperature for 5 h, then cooled to about -15ºC and held for 16 h. Toluene (5.8 L), water (11.6 L) were added sequentially, and then a solution of hydrochloric acid (37 wt%, 0.23 L, 2.8 mol) in water (1.1 L) was added, followed by line washing with water (0.12 L). The temperature of the reaction mixture was maintained at or below -9ºC throughout the addition. Remove the lower layer, add water (11.6 L) to the upper layer, stir the resulting mixture and heat to 0ºC. The lower layer was removed and the upper layer was washed twice with water (11.6 L per wash) at about 0 ° C, mixed well before sinking and each time the lower layer was removed. The remaining washed solution in the reactor was then concentrated by distilling off the solvent (8 L) under reduced pressure (210 mbar-250 mbar, 30ºC to 52ºC), then diluted with MeCN (2.9 L) and at 500 mbar Reflux for 30 min to thoroughly degas the solution. It was then cooled and held at 0ºC for 69 h (over the weekend), after which it was repeatedly depressurized and refluxed under nitrogen with any air ingress during the long hold. Then 6-chloro-7- [3- (iodomethyl) -1,5-dimethyl-pyrazol-4-yl] -1-methyl-3- (3-methylsulfonyloxypropane This solution of methyl indole-2-carboxylic acid was cooled to -14 ° C.

At the same time, 3-(((3-((acetamidothio) methyl) -1-methyl-1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ylacetate ( A mixture of intermediate 16 , 1.40 kg, 97.8 wt%, 3.42 mol) and MeOH (7.25 L) was heated to reflux and maintained at reflux under nitrogen for 30 min, and then cooled to 0ºC. Then, sodium methoxide (25 wt%, 1.6 L, 7.0 mol) was gradually added over 10 min, and then the resulting solution was warmed to 20 ° C and maintained at this temperature for 1.7 h. A portion of this solution (about 0.35 M; 6.6 L, about 2.3 mol) was gradually added to the above 6-chloro-7- [3- (iodomethyl) -1,5-dimethyl at 20 min at about -15ºC. -Pyrazol-4-yl] -1-methyl-3- (3-methylsulfonyloxypropyl) indole-2-carboxylic acid methyl ester solution. The resulting mixture was kept at -10 ° C for 1.4 h. Add another part of 3-(((1-methyl-3- (thiomethyl) -1H-pyrazol-5-yl) methyl) thio) naphthalene-1-ol disodium solution (about 0.35 M 1.5 L, approximately 0.53 mol), after which the reaction mixture was kept at -10 ° C for 18 h. The mixture was warmed to 20 ° C and removed from the reaction vessel, followed by rinsing with MeOH (0.6 L). Fill DMSO and heat to 100ºC with stirring. The intermediate solution (approximately 15 L) was then pumped back to the reactor at 100ºC over 2.8 h, followed by rinsing with DMSO (0.6 L). After maintaining the reaction mixture at 100ºC for 0.8 h, and then cooling it to slightly below 60ºC, toluene (29 L) and water (5.8 L) were added. The temperature of the mixture was adjusted to 50ºC, and then a mixture of aqueous NaOH (50 wt%, 160 g, 2.02 mol) and water (0.9 L) was added, followed by rinsing with water (0.12 L). After 10 min, the lower layer was removed. The upper layer was washed with a solution of NaCl (1.74 kg) in water (18.6 L) at 53ºC and then concentrated by distillation to remove the solvent (6 L) at 160 mbar and a jacket temperature of 85ºC to give the crude product ( 19.2 kg, 6.1 wt%, 1.7 mol) in toluene. From it with another (R _a) - methyl 6-chloro-7- (3- (hydroxymethyl) -1,5-dimethyl--1H- pyrazol-4-yl) -3- (3 -Hydroxypropyl) -1-methyl-1H-indole-2-formate ( Intermediate 9 , 1.19 kg, 98.0 wt%, 2.87 mol). Such solutions prepared according to the same procedure were combined. Kromasil ^® silica (3.0 kg, 10 μm particle size, 60 Å pore size) compression column (20 cm diameter × 22 cm length) was eluted with a mixture of toluene and ethanol (approximately 93% toluene by volume: 7 % Ethanol) to purify the combined solution by chromatography (0.84 L per portion, 51 portions). The product fractions were evaporated under reduced pressure at 50 ° C until the distillation stopped to give foamy products (91 g and 2.40 kg). The batch was dissolved in DMSO (148 g and 3.22 kg) to give ( R _a )-(+)-methyl 17-chloro-5,13,14,22-tetramethyl-28-oxa- 2,9-dithia-5,6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] tris octadecene-1 (37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35- tridecene -23- carboxylate (intermediate 17 , 239 g, 32.2 wt%, 0.112 mol and 5.63 kg, 37.9 wt%, 3.11 mol) of DMSO solution, the combined yield was 56%; m / z (ES +), [M + H] ⁺ = 686. ¹ H NMR (500 MHz, DMSO- d ₆ , 27 ºC) δ 1.97 (s, 3H), 2.16 – 2.27 (m, 1H), 2.32 – 2.42 (m, 1H), 2.89 (d, 1H), 3.08 ( d, 1H), 3.07 – 3.14 (m, 1H), 3.16 (d, 1H), 3.36 – 3.42 (m, 1H), 3.43 (d, 1H), 3.48 (s, 3H), 3.69 (s, 3H) , 3.76 (s, 3H), 3.77 – 3.83 (m, 1H), 3.84 (s, 3H), 4.13 (td, 1H), 4.22 (d, 1H), 4.29 (d, 1H), 4.77 (s, 1H ), 6.65 (d, 1H), 7.18 (d, 1H), 7.39 – 7.40 (m, 1H), 7.44 – 7.48 (m, 1H), 7.47 – 7.51 (m, 1H), 7.71 – 7.74 (m, 1H ), 7.90 (d, 1H), 8.08 – 8.12 (m, 1H). Compound 1 : ( R _a )-(+)-17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13,22 -Pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), 6 , 11,14,16,18,20,23,29,31,33,35-tridecene-23-carboxylic acid

( R _a )-(+)-methyl 17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13,22- Pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), 6, 11,14,16,18,20,23,29,31,33,35-tridecene-23-formate ( intermediate 17 , 37.9 wt% in DMSO, containing 57.3 wt% DMSO; 2.81 kg, 1.55 mol), filled DMSO (2.81 kg) and ethanol (1.68 kg) into a 20 L reactor, and heated the solution to 50 ° C with stirring. Then NaOH (50 wt% in water; 186 g, 2.33 mol) was added, followed by water (267 mL) line washing solution. After 1.5 h, add acetic acid (267 mL, 4.66 mol). The aqueous ethanol was then added (34.5 wt%; 3.0 L) , followed by addition of form _{A (R a) - (+} ) - 17- -5,13,14,22- chloro-28-oxa-2,9 four -Dithia-5,6,12,13,22-pentazaazaheptane [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 30 Octa-1 (37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formic acid monohydrate (0.8 g, 0.001 mol) crystal seed. The mixture was stirred at 50ºC for 4.5 h, and then two additional portions (4.2 and 1.3 L) of aqueous ethanol (34.5 wt%) were gradually added (over 4.1 h and 0.7 h, respectively). The slurry was cooled to 20ºC over 2 h and maintained at 20ºC for 17 h. The slurry was filtered by suction, and the filter cake was washed twice with water-based ethanol (34.5 wt%; 2.7 L per wash), and then dried in a 40ºC vacuum oven to give the form A ( R _a )-(+)-17 -Chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13,22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), 6, 11, 14, 16, 18, 20, 23, 29,31,33,35-tridecene-23-formic acid monohydrate ( compound 1 , 1.01 kg, 99.4 wt%, ≥ 99.9% ee, 94%); m / z (ES +), [M + H] ⁺ = 672. ¹ H NMR (500 MHz, DMSO- d ₆ , 27 ºC) δ 1.96 (s, 3H), 2.16 – 2.28 (m, 1H), 2.30 – 2.42 (m, 1H), 2.88 (d, 1H), 3.06 ( ddd, 1H), 3.12 (d, 1H), 3.18 (d, 1H), 3.42 (d, 1H), 3.41 – 3.48 (m, 1H), 3.51 (s, 3H), 3.72 (s, 3H), 3.75 (s, 3H), 3.84 (td, 1H), 4.09 (td, 1H), 4.25 (d, 1H), 4.28 (d, 1H), 4.75 (s, 1H), 6.67 (d, 1H), 7.13 ( d, 1H), 7.37 – 7.40 (m, 1H), 7.43 – 7.47 (m, 1H), 7.45 – 7.50 (m, 1H), 7.68 – 7.74 (m, 1H), 7.84 (d, 1H), 8.07 – 8.13 (m, 1H), 13.36 (bs, 1H).

Chiral purity analysis HPLC method details: column = ChiralPak IE-3, 3 µm 4.6 x 250 mm; temperature = 40ºC; mobile phase = 50: 50 ethanol: hexane (by volume) with 0.1% TFA, flow rate 0.8 mL / min; UV at 305 nm by the detection; injection volume = 10 μL (which can be adjusted to achieve proper detection limit); retention time R _a and S _a enantiomers were 8.5 and 11.5 min.

XRPD Form A as shown in FIG. 1, and the results are tabulated below (Table 1). [ Table 2. ] XRPD peaks for Form A

DSC analysis showed that Form A had an endothermic event of desolvation that started at about 121 ° C and reached a peak at about 158 ° C, followed by an endothermic event that started at about 181 ° C and reached a peak at about 194 ° C. TGA shows that Form A exhibits a mass loss of about 4.0% after heating from about 25ºC to about 160ºC. A representative DSC / TGA thermogram of Form A is shown in FIG. 2 .

The single crystal structure analysis proved that Form A was the monohydrate form. Crystallographic data: space group monoclinic system P 2 (1), unit cell size: a = 13.83 (3) Å, b = 7.578 (14) Å, c = 33.57 (6) Å, β = 90.23 (2) º, V = 3518 (12) Å ³ . X -ray powder diffraction ( XRPD ) analysis

XRPD analysis was performed using a Bruker D4 diffractometer, which is commercially available from Bruker AXS Inc ™ (Madison, Wisconsin). The XRPD spectrum is obtained by mounting a sample of the material (approximately 20 mg) for analysis on a single silicon crystal wafer holder (for example, a Bruker silicon zero background X-ray diffraction sample holder) and spreading the sample into a microscope slide Obtained in thin layers. The sample was rotated at 30 revolutions per minute (to improve counting statistics) and an X-ray having a wavelength of 1.5406 Angstroms (ie, about 1.54 Angstroms) was generated using a copper elongated narrow focusing tube operating at 40 kV and 40 mA. Come to shine. In θ-θ mode, the sample is exposed for 2-second increments of 0.02º (continuous scan mode) for 1 second in a 2-θ range from 2º to 40º. The execution time is 31 minutes and 41 seconds.

The XRPD 2θ value can be varied within a reasonable range, such as within a range of ± 0.2º, and the XRPD intensity may be measured when measuring substantially the same crystal form for various reasons including, for example, a preferred orientation Variety. The principles of XRPD are described in publications, such as Giacovazzo, C, et al. (1995), Fundamentals of Crystallography, Oxford University Press; Jenkins, R and Snyder, RL (1996), Introduction to X-Ray Powder Diffractometry, John Wiley & Sons, New York); and Klug, HP & Alexander, LE (1974), X-ray Diffraction Procedures [X-Ray Diffraction Program], John Wiley & Sons, New York. DSC analysis

For samples prepared according to standard methods, DSC analysis was performed using a Q SERIES ™ Q1000 DSC calorimeter available from TA INSTRUMENTS® (Newcastle, Delaware). A sample (approximately 2 mg) was weighed into an aluminum sample pan and transferred to the DSC. The instrument was purged with nitrogen at 50 mL / min and data was collected between approximately 22ºC and 300ºC using a dynamic heating rate of approximately 10ºC / minute. Thermal data is analyzed using standard software, such as Universal v.4.5A from TA INSTRUMENTS®. Thermogravimetric analysis ( TGA )

For samples prepared according to standard methods, TGA was performed using a Q SERIES ™ Q5000 thermogravimetric analyzer available from TA Instruments Instruments (Newcastle, Delaware). The sample (approximately 5 mg) was placed in an aluminum sample pan and transferred to a TGA oven. The instrument was purged with nitrogen at 50 mL / min, and data was collected between 25ºC and 300ºC using a dynamic heating rate of 10ºC / min. Thermal data is analyzed using standard software, such as Universal v.4.5A from TA INSTRUMENTS®. Example 1 : In vitro activity of compound 1

Determination of caspase activity: This line is used to measure in MOLP-8 (multiple myeloma), KMS-12-BM (multiple myeloma), MV4; 11 after 6 h treatment with Mcl-1 inhibitor (Acute myelogenous leukemia), and NCI-H23 (non-small cell lung cancer) cells to induce apoptosis. On the first day, 3000 (MOLP-8, KMS-12-BM, MV4; 11) or 1250 (NCI-H23) cells / well were seeded in 50 µL of growth medium (384-well white microplate) ( IMDM + 10% FBS + 2 mM L-Glu was used for MV4; 11, and RPMI-1640 + 10% FBS + 2 mM L-Glu was used in all other classes) and incubated overnight (37ºC, 5% C0 ₂ , 80% RH). On the next day, these cells were treated with Compound I using an ECHO acoustic liquid processor (10: 30-log serial dilution, 31.5 μm maximum concentration, 0.3% final DMSO concentration). After 6 h of incubation (37ºC, 5% C0 ₂ , 80% RH), add 25 µL of caspase-Glo 3/7 reagent (Promega) to each well, and The plates were incubated for 30 min at room temperature in the dark. Luminescence was recorded using an Infinite M200 microplate reader (Tecan) with an integration time of 100 ms. GeneData EC ₅₀ values _were calculated using analysis software and are shown in Table 2. [ Table 2. ] Results from in vitro caspase activity measurement

[ Figure 1 ] Shown as Form A ( R _a ) -17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13, 22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), Powder X-ray diffraction pattern of 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formic acid monohydrate.

[ Figure 2 ] Shown as Form A ( R _a ) -17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13, 22-pentazepine ring [27.7.1.1 ^4,7 .0 ^11,15 .0 ^16,21 .0 ^20,24 .0 ^30,35 ] 38 carbon-1 (37), 4 (38), Differential scanning thermal analysis (DSC) and thermogravimetric analysis (TGA) traces of 6,11,14,16,18,20,23,29,31,33,35-tridecene-23-formic acid monohydrate .

Claims (1)

A compound selected from: Or its salt.